31
views
0
recommends
+1 Recommend
1 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Assessment of a metabarcoding approach for the characterisation of vector-borne bacteria in canines from Bangkok, Thailand

      research-article

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Background

          Globally, bacterial vector-borne disease (VBD) exerts a large toll on dogs in terms of morbidity and mortality but nowhere is this more pronounced than in the tropics. Tropical environments permit a burgeoning diversity and abundance of ectoparasites some of which can transmit an extensive range of infectious agents, including bacteria, amongst others. Although some of these vector-borne bacteria are responsible for both animal and human diseases in the tropics, there is a scarcity of epidemiological investigation into these pathogens’ prevalence. The situation is further exacerbated by frequent canine co-infection, complicating symptomatology that regular diagnostic techniques may miss or be unable to fully characterise. Such limitations draw attention to the need to develop screening tools capable of detecting a wide range of pathogens from a host simultaneously.

          Results

          Here, we detail the employment of a next-generation sequencing (NGS) metabarcoding methodology to screen for the spectrum of bacterial VBD that are infecting semi-domesticated dogs across temple communities in Bangkok, Thailand. Our NGS detection protocol was able to find high levels of Ehrlichia canis, Mycoplasma haemocanis and Anaplasma platys infection rates as well as less common pathogens, such as “ Candidatus Mycoplasma haematoparvum”, Mycoplasma turicensis and Bartonella spp. We also compared our high-throughput approach to conventional endpoint PCR methods, demonstrating an improved detection ability for some bacterial infections, such as A. platys but a reduced ability to detect Rickettsia.

          Conclusions

          Our methodology demonstrated great strength at detecting coinfections of vector-borne bacteria and rare pathogens that are seldom screened for in canines in the tropics, highlighting its advantages over traditional diagnostics to better characterise bacterial pathogens in environments where there is a dearth of research.

          Electronic supplementary material

          The online version of this article (10.1186/s13071-019-3651-0) contains supplementary material, which is available to authorized users.

          Related collections

          Most cited references71

          • Record: found
          • Abstract: found
          • Article: not found

          Wolbachia: master manipulators of invertebrate biology.

          Wolbachia are common intracellular bacteria that are found in arthropods and nematodes. These alphaproteobacteria endosymbionts are transmitted vertically through host eggs and alter host biology in diverse ways, including the induction of reproductive manipulations, such as feminization, parthenogenesis, male killing and sperm-egg incompatibility. They can also move horizontally across species boundaries, resulting in a widespread and global distribution in diverse invertebrate hosts. Here, we review the basic biology of Wolbachia, with emphasis on recent advances in our understanding of these fascinating endosymbionts.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: found
            Is Open Access

            Optimizing methods and dodging pitfalls in microbiome research

            Research on the human microbiome has yielded numerous insights into health and disease, but also has resulted in a wealth of experimental artifacts. Here, we present suggestions for optimizing experimental design and avoiding known pitfalls, organized in the typical order in which studies are carried out. We first review best practices in experimental design and introduce common confounders such as age, diet, antibiotic use, pet ownership, longitudinal instability, and microbial sharing during cohousing in animal studies. Typically, samples will need to be stored, so we provide data on best practices for several sample types. We then discuss design and analysis of positive and negative controls, which should always be run with experimental samples. We introduce a convenient set of non-biological DNA sequences that can be useful as positive controls for high-volume analysis. Careful analysis of negative and positive controls is particularly important in studies of samples with low microbial biomass, where contamination can comprise most or all of a sample. Lastly, we summarize approaches to enhancing experimental robustness by careful control of multiple comparisons and to comparing discovery and validation cohorts. We hope the experimental tactics summarized here will help researchers in this exciting field advance their studies efficiently while avoiding errors. Electronic supplementary material The online version of this article (doi:10.1186/s40168-017-0267-5) contains supplementary material, which is available to authorized users.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found
              Is Open Access

              An inducible lentiviral guide RNA platform enables the identification of tumor-essential genes and tumor-promoting mutations in vivo.

              The CRISPR/Cas9 technology enables the introduction of genomic alterations into almost any organism; however, systems for efficient and inducible gene modification have been lacking, especially for deletion of essential genes. Here, we describe a drug-inducible small guide RNA (sgRNA) vector system allowing for ubiquitous and efficient gene deletion in murine and human cells. This system mediates the efficient, temporally controlled deletion of MCL-1, both in vitro and in vivo, in human Burkitt lymphoma cell lines that require this anti-apoptotic BCL-2 protein for sustained survival and growth. Unexpectedly, repeated induction of the same sgRNA generated similar inactivating mutations in the human Mcl-1 gene due to low mutation variability exerted by the accompanying non-homologous end-joining (NHEJ) process. Finally, we were able to generate hematopoietic cell compartment-restricted Trp53-knockout mice, leading to the identification of cancer-promoting mutants of this critical tumor suppressor.
                Bookmark

                Author and article information

                Contributors
                lghuggins@student.unimelb.edu.au
                anson.koehler@unimelb.edu.au
                ngocn4@student.unimelb.edu.au
                swilcox@wehi.edu.au
                bettina.schunack@bayer.com
                tawin.i@ku.th
                rebecca.traub@unimelb.edu.au
                Journal
                Parasit Vectors
                Parasit Vectors
                Parasites & Vectors
                BioMed Central (London )
                1756-3305
                8 August 2019
                8 August 2019
                2019
                : 12
                : 394
                Affiliations
                [1 ]ISNI 0000 0001 2179 088X, GRID grid.1008.9, Faculty of Veterinary and Agricultural Sciences, , University of Melbourne, ; Parkville, VIC 3052 Australia
                [2 ]GRID grid.444880.4, Faculty of Animal Sciences and Veterinary Medicine, , Tay Nguyen University, ; Buon Ma Thuot, Dak Lak 630000 Vietnam
                [3 ]GRID grid.1042.7, Walter and Eliza Hall Institute of Medical Research, ; Parkville, VIC 3052 Australia
                [4 ]ISNI 0000 0004 0374 4101, GRID grid.420044.6, Bayer Animal Health GmbH, ; Leverkusen, Germany
                [5 ]ISNI 0000 0001 0944 049X, GRID grid.9723.f, Faculty of Veterinary Medicine, , Kasetsart University, ; Bangkok, 10900 Thailand
                Article
                3651
                10.1186/s13071-019-3651-0
                6686542
                31395073
                e9b66313-0e33-466c-973d-7cd9ecc4a8b1
                © The Author(s) 2019

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 4 April 2019
                : 1 August 2019
                Funding
                Funded by: Australian Research Council Linkage Grant
                Award ID: LP170100187
                Award Recipient :
                Funded by: Bayer Animal Health GmbH
                Award ID: LP170100187
                Award Recipient :
                Categories
                Research
                Custom metadata
                © The Author(s) 2019

                Parasitology
                vector-borne disease,metabarcoding,prokaryotic pathogens,next-generation sequencing,16s community profiling,canines,tropics,mycoplasma,anaplasma,ehrlichia

                Comments

                Comment on this article